Selection of co-transformed Dendrobium Sonia 17 using hygromycin and green fluorescent protein
- 229 Downloads
Dendrobium Sonia 17 calluses were used for co-transformation study using particle bombardment. The bombarded transformed callus tissues were selected using half-strength MS medium containing 25 mg dm−3 hygromycin. Expression of green fluorescent protein (GFP) was observed in the callus and protocorm-like body (PLBs) tissues survived on the selection medium. The presence of green fluorescence protein (sgfp), hygromycin-B-phosphotransferase (hptII) and β-glucuronidase (uidA) genes in the transformed tissues were verified using PCR, Southern blot and dot blot analyses. Based on the results from PCR and expression of sgfp and uidA genes in the calluses and PLBs survived from hygromycin selection, we reported the co-transformation of sgfp, hptII and uidA genes into Dendrobium Sonia 17. GFP-expressing tissues were also observed in the regenerated transformed plantlets.
Additional key wordsbombardment β-glucuronidase PCR phosphotransferase Southern blot
green fluorescent protein
Murashige and Skoog
Unable to display preview. Download preview PDF.
The authors wish to thank Dr. J. Sheen (Boston, USA) for kindly providing the GFP constructs and Dr. S. Manickam (Universiti Putra Malaysia) for the pSMDFR plasmid. This research was supported by Malaysia government’s IRPA MOSTE- TOP-DOWN project # 01-02-04-T001.
- Doyle, J.J., Doyle, J.L.: Isolation of plant DNA from fresh tissue. — Focus 12: 13–15, 1990.Google Scholar